The direct role of selenocysteine in [NiFeSe] hydrogenase maturation and catalysis

Nat Chem Biol. 2017 May;13(5):544-550. doi: 10.1038/nchembio.2335. Epub 2017 Mar 20.


Hydrogenases are highly active enzymes for hydrogen production and oxidation. [NiFeSe] hydrogenases, in which selenocysteine is a ligand to the active site Ni, have high catalytic activity and a bias for H2 production. In contrast to [NiFe] hydrogenases, they display reduced H2 inhibition and are rapidly reactivated after contact with oxygen. Here we report an expression system for production of recombinant [NiFeSe] hydrogenase from Desulfovibrio vulgaris Hildenborough and study of a selenocysteine-to-cysteine variant (Sec489Cys) in which, for the first time, a [NiFeSe] hydrogenase was converted to a [NiFe] type. This modification led to severely reduced Ni incorporation, revealing the direct involvement of this residue in the maturation process. The Ni-depleted protein could be partly reconstituted to generate an enzyme showing much lower activity and inactive states characteristic of [NiFe] hydrogenases. The Ni-Sec489Cys variant shows that selenium has a crucial role in protection against oxidative damage and the high catalytic activities of the [NiFeSe] hydrogenases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biocatalysis*
  • Desulfovibrio vulgaris / enzymology*
  • Desulfovibrio vulgaris / metabolism
  • Hydrogenase / chemistry*
  • Hydrogenase / metabolism*
  • Ligands
  • Models, Molecular
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Selenocysteine / chemistry
  • Selenocysteine / metabolism*


  • Ligands
  • Recombinant Proteins
  • Selenocysteine
  • nickel-iron-selenium hydrogenase
  • Hydrogenase